A high throughput synthesis technique was developed for the combinatorial optimization of polymers imprinted with 17□-Estradiol. This incorporated a liquid handling robot for the rapid dispensing of monomers, templates, solvents and initiator into the reaction vessels of a 96-well plate. A library of 80 polymers, each ca 80mg, was prepared using functional monomers containing acid functionalities (methacrylic acid (MAA), trifluoromethylacrylic acid (TFMAA)), basic functionalities (2-vinylpyridine (2-Vpy), 4-vinylpyridine (4-Vpy), diethyl-2-aminoethylmethycrylate (DEAEMA)) or neutral functional groups (2-hydroxyethylmethacrylate (HEMA), methacrylamide (MAAM), N-vinylpyrrolidone (NVP), acetoxystyrene (AST)). Four replicas each of imprinted and nonimprinted polymers were made for each functional monomer using ethyleneglycoldimethacrylate (EDMA) as crosslinking monomer and acetonitrile as solvent. Polymer washing and exchange of incubation solutions could be effectively carried out after transfer of the polymers to solvent resistant filter plates. The binding properties of the polymers could then be rapidly assessed in the batch mode by quantifying non-bound fractions in parallel using a fluorescence monochromator plate reader. These data showed an acceptable agreement with the data from a corresponding HPLC evaluation of the rebinding solutions. In agreement with other reports, the acid functional monomers MAA and TFMAA, the basic DEAEMA and the neutral MAAM exhibited the highest imprinting factors.